Cerebellar gene expression profiles of mouse models for Rett syndrome reveal novel MeCP2 targets

<p>Abstract</p> <p>Background</p> <p>MeCP2, methyl-CpG-binding protein 2, binds to methylated cytosines at CpG dinucleotides, as well as to unmethylated DNA, and affects chromatin condensation. <it>MECP2 </it>mutations in females lead to Rett syndrome, a neurological disorder characterized by developmental stagnation and regression, loss of purposeful hand movements and speech, stereotypic hand movements, deceleration of brain growth, autonomic dysfunction and seizures. Most mutations occur <it>de novo </it>during spermatogenesis. Located at Xq28, <it>MECP2 </it>is subject to X inactivation, and affected females are mosaic. Rare hemizygous males suffer from a severe congenital encephalopathy.</p> <p>Methods</p> <p>To identify the pathways mis-regulated by MeCP2 deficiency, microarray-based global gene expression studies were carried out in cerebellum of <it>Mecp2 </it>mutant mice. We compared transcript levels in mutant/wildtype male sibs of two different MeCP2-deficient mouse models at 2, 4 and 8 weeks of age. Increased transcript levels were evaluated by real-time quantitative RT-PCR. Chromatin immunoprecipitation assays were used to document <it>in vivo </it>MeCP2 binding to promoter regions of candidate target genes.</p> <p>Results</p> <p>Of several hundred genes with altered expression levels in the mutants, twice as many were increased than decreased, and only 27 were differentially expressed at more than one time point. The number of misregulated genes was 30% lower in mice with the exon 3 deletion (<it>Mecp2</it>^tm1.1Jae) than in mice with the larger deletion (<it>Mecp2</it>^tm1.1Bird). Between the mutants, few genes overlapped at each time point. Real-time quantitative RT-PCR assays validated increased transcript levels for four genes: <it>Irak1</it>, interleukin-1 receptor-associated kinase 1; <it>Fxyd1</it>, phospholemman, associated with Na, K-ATPase;<it>Reln</it>, encoding an extracellular signaling molecule essential for neuronal lamination and synaptic plasticity; and <it>Gtl2/Meg3</it>, an imprinted maternally expressed non-translated RNA that serves as a host gene for C/D box snoRNAs and microRNAs. Chromatin immunoprecipitation assays documented <it>in vivo </it>MeCP2 binding to promoter regions of <it>Fxyd1, Reln</it>, and <it>Gtl2</it>.</p> <p>Conclusion</p> <p>Transcriptional profiling of cerebellum failed to detect significant global changes in <it>Mecp2</it>-mutant mice. Increased transcript levels of <it>Irak1, Fxyd1, Reln</it>, and <it>Gtl2 </it>may contribute to the neuronal dysfunction in MeCP2-deficient mice and individuals with Rett syndrome. Our data provide testable hypotheses for future studies of the regulatory or signaling pathways that these genes act on.</p

The development of empathy in childhood

Previous research has identified that children and adolescents, typically males, with behaviour problems have poorer empathic skills than their nonbehaviour disordered peers (e.g. De Wied, Goudena & 'Matthys, 2005). Since increased empathy is positively associated with prosocial behaviour and negatively associated with aggression (Strayer & Roberts, 2004) investigating what factors might affect child empathy might be of value in developing proactive and reactive interventions. Chapter 1 aims to review the current knowledge-base and to highlight the variety of parental factors which may affect empathy development in the typically developing child. Limitations of the research and suggestions for future research are discussed. Understanding how empathy develops in the typically developing child is important in order to understand where and why empathy development goes wrong. ! Chapter 2 presents an empirical study investigating empathy in boys with behavioural problems. This study aimed to investigate whether empathy scores were dependant on the relationship between the observer and the observed person. The findings offer some support for the prediction that empathy scores are enhanced when participants empathise with someone they have ~ positive relationship with. The thesis concludes with a reflective paper (Chapter 3) which considers the controversy between reductionism and holism in research and practice

Annual reports of the select board and other town officers Goshen, NH for the year ending December, 2015.

This is an annual report containing vital statistics for a town/city in the state of New Hampshire

Sport in der Bundeswehr:zur Geschichte, Struktur und Funktion des Militärsports in der Bundesrepublik Deutschland

Die Arbeit untersucht die Geschichte, Struktur und Funktion des Bundeswehrsports, eingebettet in den militärischen, gesellschaftlichen und politischen Kontext. Die Untersuchung setzt zur Zeit der Befreiungskriege im 19. Jahrhundert an, zeichnet die Entwicklungsstufen des Turnens und späteren Sports in Deutschland nach, wobei sich innerhalb hierbei historische Konstanten, aber auch Brüche verzeichnen lassen. Bei dem Neuaufbau der Bundeswehr in einem demokratischen System bedachte man nicht nur die gesamte Ausrichtung der Streitkräfte mit anderen Zielen, sondern auch den Sport, der nicht der körperlichen Vorbereitung zum Zwecke der Kriegführung dienen sollte. Von der Zielsetzung und Ausrichtung des Wehrmachtssportes distanziert man sich also innerhalb der Bundeswehr und belegt den jetzt durchzuführenden Sport mit anderen Werten und Normen, welche auf den Grundsätzen von Erziehung und Ausbildung zum Staatsbürger in Uniform und der Inneren Führung beruhen

Influence of concomitant coronary artery disease on surgical aortic valve replacement

Hintergrund: Die Kombination von einem chirurgischen Aortenklappenersatz und einer Koronararterienbypass-Operation ist mit einem erhöhten (Letalitäts-)Risiko im Vergleich zu einem isolierten AKE verbunden. Es ist denkbar, dass eine PCI vor der Operation das Risiko der perioperativen Letalität und Morbidität reduzieren könnte. Ziel dieser Studie war es, (1) den Einfluss einer begleitenden KHK auf die Ergebnisse beim konventionellen chirurgischen AKE zu untersuchen und (2) die Ergebnisse von Patienten, die sich einem isolierten AKE nach kürzlich erfolgter PCI unterzogen, mit denen von Patienten zu vergleichen, die sich einem kombinierten Eingriff (AKE+CABG) unterzogen. Methoden: In dieser retrospektiven Studie wurden 355 Patienten analysiert. Die Patienten wurden in vier Gruppen unterteilt (Gr. I - AKE ohne begleitende KHK; Gr. II - Patienten mit AKE und KHK ohne Interventionsbedarf; Gr. III - Patienten mit AKE und PCI innerhalb von 6 Monaten präoperativ; Gr. IV - Patienten mit kombiniertem AKE und CABG). Schließlich wurden mittels PSM 45 Patientenpaare (aus Gruppen III und IV) gebildet und analysiert. Es wurden die perioperative Letalität und Morbidität untersucht; als Endpunkt wurde die Kombination aus 30-Tages-Letalität, ischämischem Ereignis und Blutungsereignis definiert und analysiert. Ergebnisse: Im Vergleich zur Gr. I trat der kombinierte Endpunkt in Gr. II mit 34,7% doppelt so häufig auf (p=0,006), während er mit 55,6% am häufigsten in Gr. III (p<0.001) und mit 46,9% in Gr. IV (p<0.001) auftrat. Die Blutungen traten bezogen auf Gruppe I (12,1%) in allen Gruppen signifikant häufiger auf (Gr. II 22,4%, p=0,061; Gr. III 53,3%, p<0,001; Gr. IV 34,5%, p<0,001). Die Transfusion von TK und FFP war mit 57,8% (p<0,001) und 26,7% (p=0,035) am häufigsten in Gr. III erforderlich, während EK am häufigsten in Gr. IV transfundiert wurden (70,8%, p<0,001). Nach PSM trat der kombinierte Endpunkt mit 48,9% immer noch seltener in Gr. IV als in Gr. III (55,6%) – trotz hierbei überwiegend minimal-invasiv ausgeführter Operation (91,1%) - auf. Dieser Unterschied resultierte aus einer geringeren Rate an Blutungsereignissen (35,6% versus 53,3%, p=0,137), die auf der geringeren Notwendigkeit von Transfusionen basierte. Die Dauer des ITS- und KH-Aufenthaltes waren zwischen den Gruppen vergleichbar. Schlussfolgerung: Die Analysen zeigen, dass für die Ergebnisse des operativen Aortenklappenersatzes eine begleitende koronare Herzerkrankung eine entscheidende Rolle spielt. Bei Notwendigkeit zum operativen AKE ist die gleichzeitige operative Koronarrevaskularisation einer begleitenden KHK sinnvoll. Bei Patienten, bei denen ein AKE durchgeführt werden musste, führte eine interventionelle Behandlung der koronaren Herzkrankheit vor und in unmittelbarer zeitlicher Nähe zur Operation weder zu einer Reduzierung des operativen Risikos in Bezug auf den kombinierten Endpunkt noch zu einer Verkürzung des Krankenhausaufenthalts, sondern war vielmehr mit einem erhöhten Transfusionsbedarf (vor allem TK) verbunden.Objectives: The combination of surgical aortic valve replacement and coronary artery bypass grafting is associated with an increased (mortality) risk compared to isolated AS. Thus, it is thought that percutaneous coronary intervention (PCI) before surgery could reduce the risk of perioperative mortality and morbidity. The objective of this study was, (1) to investigate the influence of concomitant CAD on outcomes in conventional SAVR and, (2) to compare the outcomes of patients undergoing isolated SAVR after recent PCI and patients undergoing a combined procedure (SAVR and CABG). Methods: This retrospective, single center study analyzed 355 patients. Patients were divided into four groups (group I - SAVR without concomitant CAD; group II - patients with SAVR and CAD without need for intervention; group III - patients with SAVR and PCI within 6 months preoperatively; group IV - patients with combined SAVR and CABG). Finally, using propensity score matching, 45 pairs of patients (from groups III and IV) were formed and analyzed. Perioperative mortality and morbidity were analyzed; the combination of 30-day mortality, ischaemic event and bleeding event was defined as endpoint. Results: The combined endpoint in group II occurred with 34.7% (p=0.006) twice as high as compared to group I, while it occurred most frequently with 55.6% in group III (p<0.001) and with 46.9% in group IV (p<0.001). Bleeding related to group I (12.1%) occurred significantly more often in all groups (group II 22.4%, p=0.061; group III 53.3%, p<0.001; group IV 34.5%, p<0.001). Transfusion of platelets and fresh frozen plasma was most frequently required in group III (57.8%, p<0.001) and 26.7% (p=0.035), while packed red blood cells were most frequently transfused in group IV (70.8%, p<0.001). After PSM, the combined endpoint of 48.9% still occurred less frequently in group IV as compared to group III (55.6%) - despite predominantly being performed by minimally invasive techniques (91.1%). This difference resulted from a lower rate of bleeding events (35.6% versus 53.3%, p=0.137), which was based on the lower need for transfusions. In particular, significantly fewer transfusions of platelets were required perioperatively after SAVR and CABG (17.8% versus 57.8%, p<0.001). The length of ICU-stay and the length of hospital stay were comparable between the groups. Conclusion: The analyses show that concomitant coronary heart disease plays an important role for the outcomes of surgical aortic valve replacement. If surgical aortic valve replacement is necessary, simultaneous surgical coronary revascularisation of concomitant CAD is reasonable. In patients who required SAVR, PCI of concomitant significant coronary artery disease prior and close to surgery did neither reduce the operative risk with regards to the combined endpoint nor shortened the length of hospital stay, but was rather associated with an increased need for transfusions (especially platelet concentrates)

Neue Steuerung im Schulsystem – Risiken und Nebenwirkungen verantwortungs- und rechenschaftsorientierter Ansätze

Seit geraumer Zeit wird von einem Paradigmenwechsel in der Steuerungsphilosophie des öffentlichen Sektors gesprochen. Output- und Wettbewerbssteuerung, essenzielle Bestandteile der sog. Neuen Steuerung, werden zunehmend auch im Zusammenhang mit dem staatlichen Bildungssystem diskutiert. Daher thematisiert die vorliegende Magisterarbeit nicht-intendierte Effekte neuer Steuerungsmechanismen im Schulsystem. Der Fokus liegt dabei auf testbasierten Rechenschaftssystemen, wie sie bereits seit einigen Jahren in England und den Vereinigten Staaten von Amerika systematisch Anwendung finden. Jedoch weisen eine Vielzahl von Presseberichten, aber auch wissenschaftliche Publikationen auf negative, nicht-intendierte Wirkungen dieser Maßnahmen hin. Im zweiten Teil der Arbeit sollen daher Ergebnisse einer explorativen Studie mit Grundschullehrkräften aus Rheinland-Pfalz vorgestellt werden, in der nach Anzeichen für evtl. ähnliche Effekte im Rahmen von Vergleichsarbeiten gesucht wurde. Die übergeordnete Intention der Arbeit besteht darin, eine differenzierte Betrachtungsweise bezüglich der Möglichkeiten und Grenzen neuer Steuerungsmechanismen im Schulsystem zu unterstützen und nicht einer Diskreditierung der Lehrerprofession Vorschub zu leisten

The Influence of APOE Genotype on Neuronal Plasticity: Implications for Alzheimer's Disease and Recovery After Head Injury

Apolipoprotein E (APOE: gene, apoE: protein) is a lipid transport protein which has a well-characterised role in response to peripheral nerve injury. It has been demonstrated that the APOE epsilon4 allele of the gene is a major risk factor for Alzheimer's disease and is associated with a poor outcome after brain injury. The mechanisms underlying this are unclear but may involve a role for apoE in neuronal repair processes. This thesis investigated the role of apoE in repair processes after brain injury and determined whether this effect was APOE genotype dependent. In order to address this, models of synaptic plasticity (in vitro and in vivo) were validated. Using these models the role of apoE could be defined. The influence of APOE genotype was determined using two different lines (human promoter and GFAP promoter) of genetically modified mice (APOE knockout, human epsilon3 and epsilon4 alleles). The main studies and results are as follows: (1) Investigation of ApoE in Relation to Degeneration/ Regeneration Using a Mouse Model of Entorhinal Cortex Lesion An in vivo model of hippocampal plasticity was characterised in C57BL/6J mice (the background strain of genetically modified mice used in this thesis). A lesion of the entorhinal cortex (ECL) was induced by stereotactic injection of ibotenic acid. Specific markers (synaptophysin, GAP-43 and MAP-2) were used to define the temporal profile of degeneration and regeneration post-ECL. In the molecular layers of the dentate gyrus, synaptic decline and fibre degeneration occurred up to 28 days post-ECL, but at 90 days post-ECL, synaptogenesis and fibre sprouting were observed. Alterations in apoE paralleled degeneration and regeneration post-ECL. ApoE was upregulated within the neuropil and reactive astrocytes at day 7 post-ECL, with a further increase in the neuropil of the outer molecular layer at day 90 post- ECL. This pattern of upregulation was similar to alterations observed for apoJ. There were minimal alterations in LRP (apoE receptor) with an increase in LRP immunoreactivity on reactive astrocytes at day 7 post-ECL. Degeneration products identified by silver staining were maximal 3 days post-ECL and absent by day 90 post-ECL. This study demonstrated the utility of the entorhinal cortex lesion as a reproducible model of hippocampal plasticity. The results also supported a role for apoE in neuronal repair processes after brain injury. (2) Analysis of APOE Genotype Influence on CNS Plasticity in Transgenic Mice Expressing Human APOE epsilon3 and epsilon4 Alleles (under a human promoter sequence) Following Entorhinal Cortex Lesion The entorhinal cortex was lesioned in APOE transgenic mice, which express human APOE alleles (epsilon3, epsilon4) in astrocytes and neurons. This study assessed APOE genotype influence on the long-term response to brain injury. Specific markers (synaptophysin, GAP-43 and MAP-2) were used to define the temporal profile of degeneration and regeneration post-ECL. Initially APOEepsilon3 mice exhibited more extensive degeneration than epsilon4 mice. However, with longer recovery the APOEepsilonA mice demonstrated an impaired reparative capacity. There were no significant differences in the extent of apoE, apoJ and LRP immunoreactivity between APOEs3 and e4 mice. There was also no significant difference in the deposition and clearance of degeneration products between APOEepsilon3 and epsilon4 mice. APOEepsilon4 mice displayed a deficit in MAP-2 immunoreactivity compared to APOEepsilon3 mice that paralleled the deficit in reparative capacity. The dendrites in the molecular layer of APOEepsilon4 mice were disorganised and disarrayed. This study indicated that the epsilon4 allele was associated with impaired neuronal repair. The underlying mechanism may involve apoE isoform differences in interactions with cytoskeletal proteins. (3) Analysis of APOE Genotype Influence on CNS Plasticity in APOE Knockouts and Transgenic Mice Expressing Human APOEepsilon3 and epsilon4 Alleles (GFAP Promoter) Following Entorhinal Cortex Lesion The entorhinal cortex was lesioned in APOE knockout, epsilon3 and epsilon4 transgenic mice (expressing human APOE in astrocytes) and the influence of APOE genotype on the long-term response to brain injury was assessed. In this study it was shown that mice expressing the APOEepsilon4 allele displayed impaired neuronal sprouting capabilities compared to APOE knockout and epsilon3 mice, however there was no difference in synaptic density between APOE knockout, epsilon3 and epsilon4 mice at any time-point. Alterations in apoE and apoJ paralleled regeneration but there was no significant difference between APOE epsilon3 and epsilon4 mice. Similarly, there were no significant differences in alterations in LRP, MAPs or extent of degeneration product clearance. This study indicated that neuronal repair mechanisms were impaired in APOEepsilon4 mice compared to APOE knockout and epsilon3 mice. There were no significant differences in dendritic structure unlike that shown in the previous transgenic line and this suggests that the underlying mechanisms may not be directly related to cytoskeletal interactions in this particular line of transgenic (astrocytic expression). (Abstract shortened by ProQuest.)